Objective To establish a method for fingerprint analysis of amino acids from honey by high performance liquid chromatography ( HPLC). Methods Amino acids of honey were concentrated by 732 cation exchange resin, and then were treated by pre-column derivatization with phenyl isothiocyanate, with praline as control peak. The chromatography was performed on a Waters Symmetry C18 ( 250 mm × 4.6 mm × 5 μm) column, with acetonitrile ∶ water (4∶1) as mobile phase A and 30 mmol/L sodium acetate ∶ acetonitrile (355∶15, acetic acid adjusting pH value to be 6.5) as mobile phase B by gradient elution. The detection wave length was set at 254 nm. The flow rate was 1.0 mL/min. The column temperature was 40℃, and the injection volume was 5μL. Results Sixteen common peaks were shown in the fingerprint of 15 batches of honey samples. The similarity for 15 batches of honey samples was in the range of 0.910 ~ 0.996 . Conclusion The fingerprint detection method is simple, practical, reproducible and specific, and can provide certain reference for quality control of honey.

OBJECTIVE:To establish a method for simultaneous determination of gallic acid,chlorogenic acid,paeoniflorin and paeonol in Fule granules. METHODS:HPLC method was adopted. The determination was performed on Ultimate XB-C18 col-umn with mobile phase consisted of acetonitrile-0.05%phosphoric acid(gradient elution)at the flow rate of 1.0 mL/min. The detec-tion wavelengths were 270 nm(gallic acid,paeonol),325 nm(chlorogenic acid)and 230 nm(paeoniflorin). The column tempera-ture was 30 ℃,and sample size was 10 μL. RESULTS:The linear ranges of gallic acid,chlorogenic acid,paeoniflorin and pae-onol were 0.0762-1.524 μg(r=0.9997),0.0376-0.751 μg(r=0.9999),0.0303-0.606 μg(r=0.9997),0.0206-0.412 μg(r=0.9998),respectively. The limits of quantification were 0.353,0.276,0.421,0.540 μg/mL,and the limits of detection were 0.121,0.104,0.148,0.186μg/mL. RSDs of precision,stability and reproducibility tests were all no more than 2.04%. The recover-ies were 95.24%-100.47%(RSD=1.59%,n=9),99.49%-103.70%(RSD=2.27%,n=9),96.27%-101.09%(RSD=1.94%,n=9),95.05%-98.89%(RSD=1.22%,n=9), respectively. CONCLUSIONS:The method is simple,accurate and reproducible,and can be used for the simultaneous determination of gallic acid,chlorogenic acid,paeoniflorin and paeonol in Fule granules.

Background::Homoharringtonine (HHT) is an effective anti-inflammatory, anti-viral, and anti-tumor protein synthesis inhibitor that has been applied clinically. Here, we explored the therapeutic effects of HHT in a mouse heart transplant model.Methods::Healthy C57BL/6 mice were used to observe the toxicity of HHT in the liver, kidney, and hematology. A mouse heart transplantation model was constructed, and the potential mechanism of HHT prolonging allograft survival was evaluated using Kaplan–Meier analysis, immunostaining, and bulk RNA sequencing analysis. The HHT-T cell crosstalk was modeled ex vivo to further verify the molecular mechanism of HHT-induced regulatory T cells (Tregs) differentiation. Results::HHT inhibited the activation and proliferation of T cells and promoted their apoptosis ex vivo. Treatment of 0.5 mg/kg HHT for 10 days significantly prolonged the mean graft survival time of the allografts from 7 days to 48 days ( P <0.001) without non-immune toxicity. The allografts had long-term survival after continuous HHT treatment for 28 days. HHT significantly reduced lymphocyte infiltration in the graft, and interferon-γ-secreting CD4 + and CD8 + T cells in the spleen ( P <0.01). HHT significantly increased the number of peripheral Tregs (about 20%, P <0.001) and serum interleukin (IL)-10 levels. HHT downregulated the expression of T cell receptor (TCR) signaling pathway-related genes ( CD4, H2-Eb1, TRAT1, and CD74) and upregulated the expression of IL-10 and transforming growth factor (TGF) -β pathway-related genes and Treg signature genes ( CTLA4, Foxp3, CD74, and ICOS). HHT increased CD4 + Foxp3 + cells and Foxp3 expression ex vivo, and it enhanced the inhibitory function of inducible Tregs. Conclusions::HHT promotes Treg cell differentiation and enhances Treg suppressive function by attenuating the TCR signaling pathway and upregulating the expression of Treg signature genes and IL-10 levels, thereby promoting mouse heart allograft acceptance. These findings may have therapeutic implications for organ transplant recipients, particularly those with viral infections and malignancies, which require a more suitable anti-rejection medication.